A set of two articles recently came out in Science that directly visualize two different (and likely complementary) approaches to synapse specific delivery of gene products. Plasticity at specific synapses (input specificity — we’re restricting the discussion to the dendrites of the post-synaptic neuron) requires proteins (eg. new AMPA receptors) to get to those post-synaptic compartments and membranes. But the intructions for these new proteins are contained in the nucleus with the rest of the genome. Clearly, new proteins synthesized in the soma can’t just be sent everywhere, since only specific inputs (eg. particular dendritic spines) need these new proteins. How does this happen? Hence, the postulated synaptic tag.

Two approaches

Broadly, there are two approaches to synaptic tagging: 1) mRNA is distributed widely and translated locally at tagged locations; 2) protein products are distributed widely in the bodies of dendrites but only contact/off-load at tagged synaptic specializations. This News & Views gives a nice overview of the two papers, which find approach 1) in Aplysia cultures with sensorin mRNA and approach 2) in rat hippocampal neurons with Vesl-1S/Homer-1a protein. It amazes me that both were found pretty much simultaneously, but that might have more to do with the use of the photoconvertible Dendra2 protein than anything else.

With both approaches, we still don’t know why mRNA/protein is directed to a certain location. That is, we can visualize synaptic tagging but we don’t know what is the tag, its ontogeny, or the mechanism of tagging. But that might not be so important to understanding more about neural function. These new tools might allow us to image plasticity at many synapses at once, perhaps even in vivo. But before that, more work is needed… does the optical signal (from the Dendra fusion protein) correlate with degree of potentiation? Can we detect plasticity in the opposite direction, ie. synaptic depression, through another tag? (As a sidenote to approach 1), the use of 5′ and 3′ UTRs as a sort of molecular zipcode is also intriguing.)

Another channelrhodopsin breakthrough from Deisseroth’s lab. This time light is not required to keep the channel open. Light merely triggers opening and closing behavior. Blue-shifted light opens channels and red-shifted light closes them. This looks like another potentially powerful neurotechnology for interrogating circuits and systems.

Just the optical engineering alone here deserves mention: 28 frames per second at 62nm resolution (well below the diffraction limit of 260nm for light of the wavelength used)! STED (or stimulated emission depletion, developed in Stefan Hell’s group) is ideal for visualizing synaptic vesicles, whose small size (~50nm) has typically confined them to the domain of electron microscopists. The ability to get high-speed STED allowed the researchers to track individual vesicles and their path dynamics. They conclude that vesicle movement has both motor-driven and diffusive components (ie. a biased random walk). I’m sure with more time and more analysis there will be a lot of interesting applications for this kind of real-time vesicle tracking. Perhaps in the near future we will have single vesicle “minis” monitored at multiple sites through microscopy instead of just one or two sites electrophysiologically…

Here’s the resolution difference between STED and confocal for a single vesicle:

16384 transistors on an area of one square millimeter record the neural activity in the brain.

Hmmm, that sounds like a lot of transistors… what kind of voltage sensing resolution can a device like that provide? Well, that works out to 1.6 transistors per 10 square microns, which is arguably the relevant area for a neuron. Although these are extracellular signals, this high-resolution tool is going to have quite a large impact.

From the abstract:

We report on the recording of electrical activity in cultured hippocampal slices by a multi-transistor array (MTA) with 16384 elements. Time-resolved imaging is achieved with a resolution of 7.8 µm on an area of 1 mm2 at 2 kHz. A read-out of fewer elements allows an enhanced time resolution. Individual transistor signals are caused by local evoked field potentials. They agree with micropipette measurements in amplitude and shape. The spatial continuity of the records provides time-resolved images of evoked field potentials and allows the detection of functional correlations over large distances. As examples, fast propagating waves of presynaptic action potentials are recorded as well as patterns of excitatory postsynaptic potentials across and along cornu ammonis.

The original article (whichs seems to online in a preprint form) has excellent photos of the array (showing how it can cover a lot of a hippocampal slice), the tight correspondence between the transistor signal and a microelectrode field signal, and some cool readouts of the “whole hippocampus” with various blockers. I doubt anyone has ever been able to simultaneously do such fine scale electrophysiology on such a large portion of the mammalian brain ever before.

Treatment of striatal neurons with a D1 receptor agonist led to an increase in the dendritic staining intensity of NMDA receptor NR2B subunits. There was also an increase in the association of NR2B subunits with PSD-95 — a scaffold protein required for the assembly of NMDA receptors — and in the surface localization of NR2B-containing receptors.